Dichloroacetate increases skeletal muscle pyruvate dehydrogenase activity during acute limb ischemia

The purpose of this study was to evaluate the effects of dichloroacetate sodium (DCA), a drug that inactivates pyruvate dehydrogenase kinase (PDH-K), on pyruvate dehydrogenase (PDH) activity, lactate level, and function of skeletal muscle in an experimental model of acute limb ischemia. Thirty-two male Sprague-Dawley rats underwent right iliac artery ligation to produce hindlimb ischemia. After 2 hours of ischemia, 16 animals received intravenous DCA (15 mg/100 g body weight) and 16 control animals received an equivalent volume of normal saline. After an additional 1 hour of ischemia (total 3 hours) tibialis anterior muscle from the ischemic limb and contralateral nonischemic limb was excised, rapidly freeze-clamped with Wallenberg tongs cooled in liquid nitrogen, and stored at -70 degrees C. Muscles specimens were subsequently assayed for PDH activity and lactate level by use of spectrophotometric techniques. An additional 16 animals (DCA-treated, n = 8; control, n = 8) underwent ex-vivo gastrocnemius muscle fatigue testing with a 10 g tension preload after 3 hours of limb ischemia. In ischemic hind limbs, DCA treatment significantly (p = 0.025) increased PDH activity (19.6 +/-1.6 micromol/min/g dry weight) compared to controls (13.1 +/-1.3 micromol/min/g dry weight). DCA treatment did not increase (p = 0.13) skeletal muscle PDH activity in the nonischemic limbs (9.6 +/-1.1 micromol/min/g dry weight, controls; 13.2 +/-1.3 micromol/min/g dry weight, DCA group). In DCA-treated animals, hind limb ischemia resulted in no significant increase in muscle lactate levels compared to the nonischemic limb, while control animals demonstrated a significant (p = 0.005) elevation in lactate level in ischemic limbs compared to contralateral nonischemic limb. Ischemia induced a significant decrease in time to muscle fatigue in both DCA-treated and control animals (p = 0.002 and 0.001, respectively). Time to muscle fatigue in DCA-treated animals was increased compared to controls (2.6 +/-0.3 versus 2 +/-0.6 minutes; p < 0.05)in ischemic limbs but was not significantly different in nonischemic limbs (DCA = 3.3 +/-0.5 minutes; control = 3.1 +/-0.6 minutes). Treatment with DCA during acute limb ischemia reduced the depression of PDH activity and lactate level of skeletal muscle. Ischemic muscle function was also improved by DCA treatment. Further investigation of the potential beneficial effects of DCA treatment on muscle injury during ischemia and reperfusion is warranted.     

Reduction in spinal cord postischemic lactic acidosis and functional improvement with dichloroacetate

Pyruvate dehydrogenase complex (PDHC) is a major enzyme of glucose metabolism. Dichloroacetate (DCA) is a noncompetitive inhibitor of PDHC kinase, an enzyme that inactivates PDHC. We examined the effects of DCA on extracellular lactate and pyruvate concentration changes and spinal somatosensory evoked potentials (SSEP) in ischemic rabbit spinal cords. In the first group of 26 animals, the aorta was occluded until postsynaptic SSEP waves were completely suppressed for 10 min, a period of ischemia that causes neurologic deficits in 50% of untreated animals. DCA (25 mg/kg) was given to 13 of these animals before ischemia. In the second group of 24 animals, the aorta was occluded until the postsynaptic SSEP waves were absent for 20 min, a period of ischemia that produces paraplegia in 100% of untreated animals. DCA (25 mg/kg) was given to 16 of these animals just before the aortic occlusion was released. After occlusion, extracellular spinal lactate concentrations increased abruptly while pyruvate concentrations fell. Both lactate and pyruvate concentrations reached a plateau during the ischemic period but increased when the aortic balloon was deflated. DCA-treated animals had lower lactate and pyruvate peak concentrations during reperfusion, as well as more rapid and greater recovery of SSEP at 2 h after reperfusion. DCA did not alter spinal metabolism during the ischemia but appeared to produce a more rapid shift to glucose metabolism on reperfusion. Thus, DCA treatment resulted in better electrophysiological recovery after both moderate and severe ischemia, either by reducing lactic acidosis or by increasing the recovery rate of aerobic energy production.     

Protective effect of anisodamine on reperfusion injury of skeletal muscles in rabbit

Anisodamine is an alkaloid isolated from a Chinese plant, which was subsequently synthesized. Its chemical structure is similar to atropine. It inhibits cholinergic nerve function, improves microcirculation, and was reported to have a protective effect on reperfusion injury in various organs. We used anisodamine in a rabbit model with ischemia and reperfusion injury of hind limb muscles. We evaluated its effect on skeletal muscle cells, using transmission electron microscopy, and analyzed lipid peroxidation by measuring malondialdehyde and lactate dehydrogenase blood concentrations. We found that malondialdehyde and lactate dehydrogenase concentrations after 1 hour of reperfusion were lower in animals treated with anisodamine than in controls. Damage to membrane structures and myofilaments in muscle cells was less severe after anisodamine treatment. Our findings indicate that anisodamine protects skeletal muscles with ischemia and reperfusion injury.     

Simvastatin pretreatment reduces the severity of limb ischemia in an experimental diabetes model

OBJECTIVE: The purpose of this study was to examine the effects of simvastatin pretreatment in the setting of acute limb ischemia-reperfusion injury in an experimental diabetes model that is associated with a high risk for limb loss. METHODS: Adult male Sprague-Dawley rats were randomized into two groups. Diabetes was induced in the first group by intravenous streptozotocin injection. The second group served as the nondiabetic group. Eight weeks after the streptozotocin injection, half of the rats in the diabetic and the nondiabetic groups were further randomized to receive either intraperitoneal simvastatin (1 mg/kg per day) or saline treatment for 6 weeks. Bilateral hind-limb ischemia was induced for 4 hours by the tourniquet method. After 24 hours of reperfusion, tissue samples were collected from the gastrocnemius and anterior tibial muscles bilaterally for measurement of muscle edema, percentage of necrosis, and malondialdehyde (MDA), glutathione, and myeloperoxidase (MPO) levels. RESULTS: Ischemic injury was more prominent in diabetic animals. The diabetic animals with limb ischemia exhibited a 7% increase in tissue edema, a 47% increase in muscle necrosis and MPO level, and a 15% reduction in glutathione levels compared with the nondiabetic animals (P < .05). Simvastatin treatment with 1 mg/kg for 6 weeks reduced the ischemic injury. Simvastatin pretreatment led to a 71% reduction in muscle necrosis in diabetic animals (P < .001). The protective effects of simvastatin pretreatment also correlated with a 23% improvement in tissue edema, a 75% reduction in tissue myeloperoxidase content, and a 71% increase in glutathione levels in diabetic animals (P < .01). Furthermore, skeletal muscle injury, characterized by tissue edema and leucosequestration, was significantly less severe with simvastatin pretreatment compared with the nondiabetic animals (P < .01). CONCLUSION: Simvastatin pretreatment reduced limb ischemia-reperfusion injury in diabetic and nondiabetic animals. We conclude that simvastatin pretreatment may be a potential therapeutic intervention for skeletal muscle ischemia-reperfusion injury in the clinical setting.     

Metabolic alterations of skeletal muscle tissue after prolonged acute ischemia and reperfusion.

Acute tissue ischemia is usually followed by considerable disturbances of cellular metabolism that often lead to cell death. Reperfusion improves cellular function by withdrawing the toxic products of ischemia and providing energy sources, although sometimes it worsens it. The purpose of this experimental work is to study the metabolic disturbances in skeletal muscle tissue of canines after prolonged acute ischemia in relation to the values of certain substances (ATP, lactate, pyruvate, the ratio of lactate to pyruvate [L/P], and glucose) and whether the alterations in the values of these substances could be used as prognostic indices of the magnitude of tissue damage and the possibility of inverting it. We used 15 mongrel dogs. Complete acute ischemia was induced in the right lower limb lasting 12 h. Reperfusion also lasted for 12 h. The left lower limb was used as reference value. Before the beginning of ischemia, at (1/2), 1, 6, and 12 h after the induction of ischemia, and at (1/2), 1, 6, and 12 h after the restoration of circulation, blood samples, and tissue biopsies were obtained from the healthy and the experimental limb for the measurement of ATP, lactate, pyruvate, the ratio of L/P, and glucose. From the statistical analysis of the values of the controlled parameters the following were concluded: The changes in ATP, lactate, pyruvate, and the L/P ratio in the venous blood of the experimental limb and in the intracellular space of the suffering skeletal muscle could be used as indices to evaluate ischemic injury to the skeletal muscles, the course of its development, and the possibility of reversal after reperfusion.     

Pharmacological reversal of abnormal glucose regulation, BCAA utilization, and muscle catabolism in sepsis by dichloroacetate

Sepsis has been shown to decrease skeletal muscle glucose oxidation by inhibiting the pyruvate dehydrogenase activity (PDHa) and to increase proteolysis and use of branched-chain amino acids (BCAA). The effects of dichloroacetate (DCA), which reverses PDHa inhibition, were studied in skeletal muscle from a septic (S) rat model of intra-abdominal abscess (E. coli + B. fragilis) and compared to control (C) and sterile inflammatory abscess (I) animals. In one set of S, I, and C animals, DCA (1 mmol/kg) was injected intraperitoneally at 0, 30, and 60 min. Septic, but not I, rats had a twofold increase in skeletal muscle lactate concentrations over C, but no changes in pyruvate. After DCA, both lactate and pyruvate were reduced (p less than 0.001) to same level in S, I, and C. Skeletal muscle alanine was increased in S compared to I or C, but after DCA was reduced threefold in C, S, and I (p less than 0.001) suggesting that alanine synthesis may be impaired due to decreased pyruvate availability. Like alanine, skeletal muscle BCAA were increased in S compared to C, but not altered in I. Following DCA, BCAA levels in muscle from S were reduced (p less than 0.001) to values seen in C or I. Muscle phenylalanine content was significantly elevated in S (p less than 0.05) compared to C or I, but was reduced (p less than 0.05) after DCA in S but not in C or I. Decreased muscle phenylalanine associated with lowered BCAA suggests DCA may decrease septic muscle protein catabolism and/or enhance protein synthesis. Coupled with an increased PDHa and reduced lactate levels, this suggests that DCA may reverse the excess muscle catabolism and BCAA dependence of sepsis by increasing glucose and lactate oxidation and may be a useful therapeutic modality.     

Effect of starvation on the local and systemic metabolic effects of the lambda-carrageenan wound

The contribution that starvation makes to the altered glucose metabolism in injured rats was evaluated. Food intake, weight change, nitrogen balance, and muscle tissue concentrations of glycogen, glucose, and the glycolytic intermediates were determined in these animals. This study concluded that the wounded and pair fed control groups presented adequately represent the metabolic states associated with injury and semistarvation in experimental animals, decreased food intake plays a major role in the weight loss and nitrogen balance in this wound model, wounding overrides two of the controlling steps of glycolysis (hexokinase and phosphofructokinase) in skeletal muscle during starvation, the finding of similar pyruvate dehydrogenase activity after wounding and starvation as demonstrated by tissue lactate to pyruvate ratios and lactate and pyruvate concentrations suggest that lactate production in wounded tissue may not be simply a manifestation of an altered redox state secondary to anaerobic conditions.     

An isolated skeletal muscle model suitable for acute ischemia studies

A modified isolated canine gracilis model of acute complete muscle ischemia was developed and then tested metabolically and histologically in 25 animals to assess its validity. In each dog, both gracili were isolated on their major vascular pedicles. One muscle underwent ischemia and reperfusion by placing and removing microvascular clips on the artery and vein. The other gracilis muscle was used as a control. Total muscle blood flow measurements, blood samples, and muscle biopsies were taken every other hour for up to 11 hr after preparation. The fiber-type profile of the gracilis was determined bilaterally using a myosin ATPase stain (n = 10). The results verified these hypotheses: after surgical preparation, the right and left muscles in the same dog are equivalent metabolically, after a 2-hr stabilization period, gracilis blood flow, oxygen and glucose uptake, lactate release, and tissue glycogen, lactate, phosphocreatine, and ATP levels remain within normal limits and unchanged for the next 9 hr, the surgical isolation of the gracilis muscle on a single vascular pedicle does not result in significant metabolic changes, in this model, a 2-hr ischemia is reversible, but a 7-hr ischemia results in irreversible ischemic injury. As well, fiber-type profile, muscle blood flow, and metabolic parameters can very significantly among animals supporting the necessity of a contralateral control. Therefore, this modified gracilis muscle model with its contralateral muscle as a control is suitable for acute skeletal muscle ischemia experiments of at least 9-hr duration.     

Local but not systemic capillary lactate is a reperfusion biomarker in experimental acute limb ischaemia

IntroductionSystemic capillary lactate, an end product of cellular anaerobic metabolism, has not established credibility in monitoring limb reperfusion. We assessed, in mice, whether local capillary lactate, arising from the reperfused limb, might be a relevant biomarker of reperfusion.ReportSystemic and local capillary lactate were sampled in the non-ischaemic and in the ischaemic limb. Only local lactate concentrations significantly increased after 2 h of ischaemia and decreased after reperfusion.DiscussionLocal, but not systemic, capillary lactate appeared as a potential reperfusion biomarker in this experimental acute limb ischaemia model.     

Effect of extended cold ischemia time on glucose metabolism in liver grafts: experimental study in pigs

Background/Purpose Recovery of normal carbohydrate metabolism in the liver after transplantation is highly important. The aim of the present study was to evaluate how short and long cold ischemia (CI) time followed by warm ischemia (WI) impact intrahepatic glucose metabolism in a pig liver transplantation model. Methods Twenty-six animals were divided into two transplantation groups: group I with a liver ischemia time of 5 h (n = 6), and group II with 15 h of liver ischemia (n = 7). Intrahepatic microdialysis samples were collected throughout the experiment at 20-min intervals, during the donor operation, cold preservation, liver implantation, and liver reperfusion in the recipient. Glucose, lactate, and pyruvate concentrations were analyzed and the lactate/pyruvate ratio (L/Pr) was calculated. Result There were no changes in glucose levels during CI. However, during WI, glucose and lactate increased and the increase was significantly higher in the group with longer CI (P < 0.01). The L/Pr increased at the beginning of CI but accelerated to increase during WI in both groups, with significantly prolonged and higher levels in the group with longer CI (P < 0.01). Conclusions Extended CI results in increased intrahepatic glycogenolysis, delayed restoration of aerobic glycolysis, and prolonged anaerobic glycolysis shortly after reperfusion. Improvements in glycogen protection and faster restoration of aerobic metabolism during preservation and reperfusion time seem to be necessary in order to improve liver preservation protocols per se.     

The value and limitation of iloprost infusion in decreasing skeletal muscle necrosis.

Iloprost has been shown to minimize skeletal muscle necrosis when administered before the onset of ischemia in animal experiments, possibly by preventing neutrophil activation. Since patients with acute limb ischemia are seen after the process has begun, we investigated whether iloprost can be protective when given only during reperfusion. After anesthesia, 18 adult mongrel dogs underwent a standard isolated gracilis muscle preparation. In six control animals (group I) the gracilis muscle was subjected to 6 hours of ischemia followed by 48 hours of reperfusion. Group II animals (n = 6) received intravenous infusion of iloprost at a dose of 0.45 microgram/kg/hr beginning 1 hour before the onset of muscle ischemia and throughout the experiment (6 hours of ischemia and 1 hour of reperfusion). In addition to the continuous infusion, they received 0.45 microgram/kg intravenous boluses of iloprost 10 minutes before the induction of ischemia and 10 minutes before reperfusion. Group III animals (n = 6) had a similar ischemic interval, but were given a bolus of iloprost of 0.45 microgram/kg at end ischemia followed by continuous infusion of 0.45 microgram/kg/hr for 48 hours during reperfusion. Muscle biopsies were obtained at baseline and after 1 hour of reperfusion in all groups. Additional biopsies were obtained at 48 hours of reperfusion in groups I and III. Myeloperoxidase activity, a marker of neutrophil activation, was measured in all muscle biopsies. At the end of reperfusion, the gracilis muscle was harvested in all animals and weighed. Muscle necrosis was estimated by serial transection, nitroblue tetrazolium histochemical staining followed by computerized planimetry.(ABSTRACT TRUNCATED AT 250 WORDS)     

大鼠后肢急性缺血模型的构建及评估

目的：探讨大鼠后肢急性缺血后血流、血压的动态变化及肌肉组织的变化,为进行后续干预实验摸索条件。方法：取Lewis大鼠10只采用切除左后肢股动脉及其分支至膝关节附近的方法制备急性后肢缺血模型,分别于术后2,7,14,28,42,49 d对患侧和健侧肢体进行血流、血压及血管造影分析。取材后行后肢肌肉HE常规染色,观察病理变化。结果：动物术后均成活,但部分出现后肢跛行(n=2),紫绀（n=2）,苍白(n=3),肌肉的萎缩(n=1)。急性缺血下肢部分肌肉坏死,大量炎性细胞浸润;后期出现纤维化。患肢血流在术后7 d明显下降,术后28 d下降至最低点,术后49 d基本恢复正常（正常肢体的90%）。患肢血压在术后2 d明显下降,14 d下降至最低点,后逐渐恢复,至49 d基本恢复正常。结论：结扎并切除大鼠后肢股动脉及分支可以成功制备下肢缺血模型,但血流及血压在术后42~49 d基本恢复正常,术后14~28 d是缺血最严重的阶段,适于进行各种干预实验研究。     

A comparison of acute mouse hindlimb injuries between tourniquet- and femoral artery ligation-induced ischemia-reperfusion

The tourniquet or femoral artery ligation is widely used to stop extremity hemorrhage or create a bloodless operating field in the combat scenario and civilian setting. However, these procedures with subsequent reperfusion also induce ischemia-reperfusion (IR) injuries. To fully evaluate animal models of limb IR injuries, we compared tourniquet- and femoral artery ligation-induced IR injuries in the hindlimb of mice. In C57/BL6 mice, 3 h of unilateral hindlimb ischemia was induced by placement of a rubber band at the hip joint or a surgical ligation of the femoral artery. The tourniquet or femoral artery ligation was then released, allowing for 24 h of reperfusion. Compared to the femoral artery ligation/IR, the tourniquet/IR induced more severe skeletal muscle damage, including muscle necrosis and interruption of muscle fibers. There was no gastrocnemius muscle contraction in tourniquet/IR, while femoral artery ligation/IR markedly weakened gastrocnemius muscle contraction. Motor nerve terminals disappeared, and endplate potentials (EPPs) were undetectable in tourniquet/IR, whereas femoral artery ligation/IR only induced mild impairment of motor nerve terminals and decreased the amplitude of EPPs. Additionally, western blot data showed that proinflammatory cytokine levels (IL-1β and TNF-α) were higher in the tourniquet/IR than that in femoral artery ligation/IR. Moreover, tourniquet/IR caused significant tissue edema and dilation of lymphatic vessels in the hindlimb, compared to femoral artery ligation/IR. The above data demonstrated that tourniquet/IR-induced acute hindlimb injuries are more severe than those induced by femoral artery ligation/IR. This suggests that future investigators should determine which hindlimb IR model (tourniquet/IR or femoral artery ligation/IR) is optimal depending on the purpose of their study.     

肢体缺血再灌注损伤的新型动物模型制作

[目的]用充气止血带制作肢体缺血再灌注损伤的新型动物模型,研究其对周围神经和骨骼肌损伤的影响.[方法]选择健康新西兰大白兔6个月龄,30只,体重(3.5 ±0.3) kg,雌雄不限,在家兔左侧后肢环扎充气止血带,于不同时间点松开,造成左侧后肢缺血再灌注损伤的模型.随机分为3组,每组10只.A组:对照组,B组:缺血2h,C组:缺血4h.对照组不扎充气止血带,第1、2、3、4、5、6h检测肢体的神经电生理学指标,B组、C组于再灌注(松开止血带,血供恢复后)的1、2、3、4、5、6h检测肢体的神经电生理学指标,A组于第6h观察骨骼肌的形态,B、C组于再灌注(松开止血带,血供恢复后)的第6h观察骨骼肌的形态,每组于术后第5d评估左侧后肢的行走功能.[结果]随着缺血后再灌注时间的延长,B、C和A组相比较,周围神经的潜伏期延长、波幅降低,传导速度降低,三组之间的潜伏期、波幅、传导速度差异均有统计学意义(P＜0.05),光镜观察骨骼肌可见(B、C组):横纹紊乱、肌纤维断裂、间质血管扩张充血、大量中性粒细胞浸润.[结论]经过缺血期和再灌注损伤的交互作用后,肢体的功能性损伤进一步加重,出现了不可逆的病损.该模型制作对动物的损伤较小,更贴近临床.     

Sevoflurane but not propofol increases interstitial glycolysis metabolites availability during tourniquet-induced ischaemia-reperfusion

BACKGROUND: Ischaemia/reperfusion (I/R) is one of the main pathophysiological phenomena involved in the anaesthetic practice. The authors hypothesized that anaesthetic regimens can influence skeletal muscle tolerance to tourniquet-induced I/R that should be reflected by the interstitial metabolite levels of anaerobic glycolysis. METHODS: Microdialysis probes were implanted in three groups of 10 patients each receiving either sevoflurane (SEVO), propofol (PRO), or spinal (SA) anaesthesia (for induction and maintenance). SA group was considered as a control group. Interstitial fluid was obtained during tourniquet-induced I/R and was analysed for interstitial glucose, lactate, pyruvate, and glycerol. RESULTS: The microdialysis flow rate was 0.5 microl min(-1). Compared with the control group, the SEVO group had a higher level of both lactate and pyruvate and an increase in glucose during ischaemia. In contrast, the PRO group had a lower level of pyruvate, resulting in a significant higher increase (eight times from baseline) of the lactate pyruvate ratio. Glucose level remained low in this group. During reperfusion, lactate, pyruvate, and glucose remained at a significantly higher level in the SEVO group. In the PRO group, there was no difference in lactate, pyruvate, and glucose levels compared with the control group. The interstitial level of glycerol exhibits only few and comparable changes during I/R between the groups. CONCLUSIONS: Our results indicate that there is a better availability of interstitial glycolysis metabolites (glucose, lactate, and pyruvate) in the skeletal muscle during ischaemia and reperfusion after sevoflurane exposure than after propofol, suggesting a potential preconditioning effect of sevoflurane on tourniquet-induced skeletal muscle I/R.     

Acute dichloroacetate administration increases skeletal muscle free glutamine concentrations after burn injury.

OBJECTIVE: To investigate the hypothesis that the stimulation of pyruvate oxidation by dichloroacetate (DCA) administration would increase the level of intramuscular glutamine in severely burned patients. SUMMARY BACKGROUND DATA: The level of intramuscular glutamine decreases in response to severe injury, and the rate of intramuscular glycolysis and pyruvate oxidation is elevated. Intramuscular glutamine concentrations have been correlated to muscle protein synthesis. METHODS: Six studies were conducted on five patients with burns >40% total body surface area. Patients were studied in the fed state during an 8-hour stable isotope infusion. After 5 hours, DCA (30 mg/kg) was administered for 30 minutes. RESULTS: Analysis of muscle biopsy samples taken at 5 and 8 hours of the study revealed a 32% increase in intracellular glutamine levels after DCA administration. Increased intracellular glutamine concentrations did not affect skeletal muscle protein synthesis as determined by a three-pool arteriovenous model or by the direct incorporation of isotope into skeletal muscle protein. DCA administration resulted in a decrease in plasma lactate but no change in alanine de novo synthesis or intracellular concentration. CONCLUSIONS: These results suggest that acute DCA administration can increase intramuscular glutamine concentration, but that this acute elevation does not affect muscle protein metabolism.     

Efficacy of leukocyte-depleted terminal blood cardioplegia in congenital heart disease.

OBJECTIVE: Activated leukocytes and their products play a significant role in reperfusion injury and cause microvascular occlusion--the 'no-reflow phenomenon'--which decreases coronary blood flow after the release of the aortic cross-clamp during open-heart surgery. This study was designed to evaluate the efficacy of leukocyte-depleted terminal blood cardioplegia in patients with right ventricular pressure or volume overloaded congenital heart disease undergoing intracardiac repair. SUBJECTS AND METHODS: Fourteen infants and children undergoing intracardiac repair for congenital heart defects were the subjects in this study. Leukocyte-depleted terminal blood cardioplegia was employed in seven patients (Group I). For the control, terminal blood cardioplegia with leukocytes was employed in seven patients (Group II). In order to evaluate the myocardial anaerobic metabolism, the arteriovenous difference in the pyruvate and lactate levels was compared between both groups. Blood samples were taken simultaneously from both the arterial limb of the bypass circuit and the coronary sinus immediately after, and at 5, 10, and at 20 minutes after, the release of the aortic cross-clamp. RESULTS: The coronary arteriovenous difference in the pyruvate and lactate levels demonstrated that the myocardial anaerobic metabolism in Group II was significantly higher than in Group I. The delta excess lactate demonstrated that myocardial hypoxia in Group II persisted significantly longer than in Group I. There was a tendency for the creatine kinase-MB level at 6 hr after surgery to be lower in Group I than in Group II. CONCLUSION: Leukocyte-depleted terminal blood cardioplegia may reduce the myocardial anaerobic metabolism in patients with right ventricular pressure or volume overloaded congenital heart disease undergoing intracardiac repair.     

Recovery of neuromuscular function during reperfusion of the ischemic extremity: effect of mannitol and superoxide dismutase

The ability of mannitol and superoxide dismutase (SOD) to improve the recovery of peripheral nerve and skeletal muscle function and to influence metabolism during reperfusion after 4 hours of complete ischemia was investigated in an autoperfused canine hind-limb model. Study groups included control subjects (n = 7), subjects given 5000 units/kg of SOD intra-arterial bolus immediately before reperfusion and 10,000 units/kg infusion during first hour of reperfusion (n = 7), and subjects given 150 mg/kg isosmolar mannitol intra-arterial bolus before reperfusion and 1 gm/kg intravenous infusion during the first hour of reperfusion. Function was evaluated by determining isometric twitch and tetanic contractile force of paw dorsiflexion by stimulating the peroneal nerve or the anterior tibial muscle. Metabolic responses (oxygen consumption and lactate clearance) and blood flow were not influenced by either treatment protocol. However, mannitol significantly reduced muscle damage and significantly improved neuromuscular contractile function compared to control and SOD treatment regimens.     

Monitoring of porcine myocardial ischemia and reperfusion by intravasal microdialysis

OBJECTIVE: A novel application of microdialysis is studied where myocardial metabolism is semi-continuously monitored in the myocardial venous outflow during ischemia and reperfusion. DESIGN: Microdialysis catheters were introduced into the great cardiac vein, the pulmonary artery, and the right external jugular vein in 20 anesthetized pigs. The left anterior descending artery was occluded in four separate groups of pigs for 0, 10, 15, and 60 min, respectively. Ischemia was followed by 120 min of reperfusion. Microdialysis samples were collected every 10-20 min and analyzed for lactate, pyruvate, glycerol, glutamate, and glucose. RESULTS: Myocardial infarction was observed after 15 min of ischemia. Metabolic changes were observed only in the great cardiac vein. Lactate increased early during ischemia. After 60 min of ischemia an increase of the lactate/pyruvate ratio and glutamate was observed. Glycerol was progressively released during prolonged ischemia. Myocardial infarction resulted in an additional release of glycerol early in reperfusion. CONCLUSION: Intravasal microdialysis is a semi-continuous method to monitor myocardial metabolism and tissue damage during ischemia and reperfusion.     

Comparison of histopathologic effects of carnitine and ascorbic acid on reperfusion injury.

OBJECTIVE: Reperfusion injury can be seen after acute arterial occlusion, acute myocardial infarctus and during open heart surgery and vascular surgery. Protective effects of ascorbic acid and carnitine on reperfusion damage were tested and compared using histopathologic examination on ischemia model in the rabbit hind limb. METHODS: Four groups (each containing ten animals) were used. In group I (G1), only anesthesia was administered and a biopsy was taken from the soleus muscle after 6 h. In group II (G2), group III (G3), and group IV (G4), after induction of anesthesia, arterial blood circulation of right posterior extremity was blocked by a tourniquet proximally. After four hours of ischemia, just before releasing of tourniquet, physiologic saline solution, sodium ascorbate (Redoxan) and L-carnitine (Carnitine) were administered intravenously to G2, G3 and G4, respectively. Following 2 h of reperfusion, biopsies were taken from soleus muscles. All of the biopsy slides were observed under the light microscope from the aspect of six different histopathologic criteria (loss of striation, nuclear centralisation, formation of ring and/or splitting, changing on diameters of muscle fibers, necrosis and minimal fibrosis) of ischemic muscle. RESULTS: Ischemic change criteria were seen less frequency in both vitamin C and carnitine groups compared to the control and placebo groups. However, this protective effect was statistically significant only for the aspect of segmental necrosis, centralization of nuclei and diameter change parameters in G3 and in G4. When G3 and G4 were compared, the differences on protective effects were significant only from the aspect of fibrosis (P<0.001) and changing on diameter of the fibers (P<0.001). CONCLUSIONS: Both sodium ascorbate and carnitine are effective on reducing the reperfusion injury in skeletal muscle. But when we compared these two agents to each other, we found that carnitine seems a little more protective on our experimental model.